The effects of inherited NUDT15 polymorphisms on thiopurine active metabolites in Japanese children with acute lymphoblastic leukemia

Abstract

Thiopurines [e.g. mercaptopurine (MP)] are widely used as chemotherapeutic agents in the treatment of pediatric acute lymphoblastic leukemia with dose-limiting hematopoietic toxicity. Recently, germline variants in NUDT15 have been identified as a major genetic cause for MP-related bone marrow suppression, and there is increasing interest in the clinical implementation of NUDT15 genotype-guided MP dose individualization. Therefore, we sought to evaluate the effects of NUDT15 on thiopurine metabolism and identify pharmacologic markers to inform NUDT15 genotype-guided MP dosing. In 55 Japanese children with acute lymphoblastic leukemia, we simultaneously measured both thioguanine nucleotides (TGN) in red blood cells and DNA-incorporated thioguanine (DNA-TG) in white blood cells. TGN levels were significantly lower in patients with NUDT15 deficiency, likely because of toxicity-related MP dose reduction. In contrast, when exposed to the same dose of MP, DNA-TG accumulated more efficiently in vivo with increasing number of risk alleles in NUDT15 (P=4.0×10). Cytosolic TGN and nuclear DNA-TG were correlated positively with each other across genotype groups (P=6.5×10), but the ratio of DNA-TG to TGN was significantly higher in NUDT15-deficient patients (P=3.6×10), consistent with excessive MP activation. In conclusion, our results suggest that DNA-TG is a more relevant MP metabolite than TGN to inform NUDT15 genotype-guided dose adjustments.

Fig. 1

Association of erythrocyte TGN with mononucleated cells DNA-TG. Cytosolic TGN and nuclear DNA-TG levels were highly correlated with each other, consistent with the fact that TGN is the precursor metabolite of DNA-TG during thiopurine activation (P=6.5 × 10⁻⁴, R² = 0.16). However, the regression slope of DNA-TG over TGN was significantly higher for patients with the intermediate-activity NUDT15 diplotype (the green line) than those with the normal-activity NUDT15 diplotype (the black line).

Fig. 2

Association of DNATG/TGN with NUDT15 genotypes. The ratio of DNA-TG to TGN (i.e., the percent of TGN converted to DNA-TG) was significantly higher in NUDT15 deficient patients (P = 3.6 × 10⁻⁹), suggesting that the lack of functional NUDT15 directly resulted in more robust conversion of TGN to DNA-TG, and therefore elevated risk of toxicity.